New method for labeling and autoradiographic localization of androgen receptors

Journal Article · Wed Jul 01 00:00:00 EDT 1987 · J. Histochem. Cytochem.; (United States)

DOI:https://doi.org/10.1177/35.7.3495565· OSTI ID:6332862

Peters, C A; Barrack, E R

We have used a novel receptor labeling and autoradiographic technique to localize androgen receptors in the intact rat ventral prostate at the morphological level. Frozen slide-mounted prostate tissue sections (10 micron thick) were incubated with increasing concentrations of (/sup 3/H)-R1881 in the absence and presence of excess unlabeled R1881. Tissue sections labeled in this way were subjected to concurrent biochemical and autoradiographic analysis. After incubation and washing to remove free (/sup 3/H)-steroid, some of the sections were wiped from the slides for scintillation counting in order to characterize and quantitate (/sup 3/H)-R1881 binding. Androgen receptors could indeed be labeled in slide-mounted tissue sections, and specific (/sup 3/H)-R1881 binding to these receptors was high-affinity (Kd = 1 nM), saturable, and androgen-specific. All cellular androgen receptors appear to be retained, because receptor content in sections was comparable to the sum of receptors in subcellular fractions of homogenized tissue. Replicate labeled slide-mounted tissue sections were dried rapidly, apposed to dry emulsion-coated coverslips, and exposed in the dark for autoradiography. Silver grains were counted over nuclei or cytoplasm of epithelium or stroma to evaluate specific androgen receptor location. Autoradiographic analysis demonstrated androgen receptor localization almost exclusively in the epithelial nuclei, with little or none in the stroma. We discuss here the unique features and advantages of labeling androgen receptors in slide-mounted frozen tissue sections for autoradiographic localization.

• Research Organization: Johns Hopkins Univ. School of Medicine, Baltimore, MD
• OSTI ID: 6332862
• Journal Information: J. Histochem. Cytochem.; (United States), Journal Name: J. Histochem. Cytochem.; (United States) Vol. 7; ISSN JHCYA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AFFINITY
ANDROGENS
ANDROSTANES
ANIMAL TISSUES
ANIMALS
AUTORADIOGRAPHY
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL LOCALIZATION
BODY
CELL CONSTITUENTS
CELL NUCLEI
COUNTING TECHNIQUES
EPITHELIUM
GLANDS
HORMONES
KINETICS
LABELLED COMPOUNDS
LABELLING
MALE GENITALS
MAMMALS
MEMBRANE PROTEINS
ORGANIC COMPOUNDS
ORGANS
PROSTATE
PROTEINS
RATS
REACTION KINETICS
RECEPTORS
RODENTS
SCINTILLATION COUNTING
STEROID HORMONES
STEROIDS
TISSUES
TRITIUM COMPOUNDS
VERTEBRATES